Identification of Inhibitors of the Antibiotic-Resistance Target New Delhi Metallo-尾-lactamase 1 by both Nanoelectrospray Ionization Mass Spectrometry and Ultrafiltration Liquid Chromatography/Mass Sp

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• 作者：Xin Chen ; Lixin Li ; Shuai Chen ; Yintong Xu ; Qiang Xia ; Yu Guo ; Xiang Liu ; Yanting Tang ; Tanjie Zhang ; Yue Chen ; Cheng Yang ; Wenqing Shui
• 刊名：Analytical Chemistry
• 出版年：2013
• 出版时间：August 20, 2013
• 年：2013
• 卷：85
• 期：16
• 页码：7957-7965
• 全文大小：504K
• 年卷期：v.85,no.16(August 20, 2013)
• ISSN：1520-6882

文摘

Mass spectrometry-based platforms have gained increasing success in discovery of ligands bound to therapeutic targets as drug candidates. We established both a nanoelectrospray ionization mass spectrometry (nanoESI-MS) assay and an ultrafiltration liquid chromatography/mass spectrometry (LC/MS) assay to identify new ligands for New Delhi metallo-尾-lactamase 1 (NDM-1), responsible for worldwide antibiotic resistance. To alleviate nonspecific binding of hydrophobic compounds and eliminate false positives typically encountered in the indirect LC/MS-based assay, we introduced a blocking protein in the control, which remarkably enhances the selectivity and accuracy of the indirect approach. Side-by-side comparison of the two MS-based approaches for the first time further reveals unique advantages of the indirect approach, including better reproducibility and tolerance of interference. Moreover, the success of fishing out a potent ligand from a mixture of small-molecule fragments demonstrates great potential of the indirect LC/MS-based approach for constructing a robust screening platform against combinatorial libraries or natural product extracts. More importantly, by combining the results of MS-based analyses, enzymatic activity assay, competition experiments, and structural simulation, we discovered a new compound as a promising drug candidate targeting NDM-1.